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The effect of treatment and co\u2010culture on production of embryos of the preferred sex from unsorted sperm was also studied. Sperm from five breeding bulls was used for fertilization of mature oocytes as follows<jats:bold>:<\/jats:bold> Experiment 1, sorted and unsorted sperm (bulls A\u2010E) treated only with heparin in standard co\u2010cultures; Experiment 2, sorted sperm (bulls A\u2010E) treated with heparin\u2010PHE (penicillamine, hypotaurine, and epinephrine) or heparin\u2010caffeine in drop co\u2010cultures; and Experiment 3, unsorted sperm (bull E) treated with either heparin\u2010PHE or heparin\u2010caffeine in both standard and drop co\u2010cultures. In all bulls, treatment with heparin resulted in significantly (<jats:italic>p<\/jats:italic>\u00a0&lt;\u00a0.05) reduced cleavage and blastocyst rates from sorted sperm, as compared with those from unsorted sperm. In bulls A, B, D and E, treatment of sorted sperm with heparin\u2010PHE in drops significantly increased the blastocyst rate (<jats:italic>p<\/jats:italic>\u00a0&lt;\u00a0.05). In unsorted sperm of bull E, heparin\u2010PHE treatment in drops resulted in the XX\/XY sex ratio inverse to that obtained by heparin\u2010caffeine treatment in standard co\u2010cultures (32.3%\/67.7% and 66.7%\/33.3%, respectively). In conclusion, the treatment of sorted sperm with heparin\u2010PHE in modified drop co\u2010cultures can be recommended for production of i<jats:italic>n vitro<\/jats:italic> sexed embryos. The use of unsorted sperm for production of embryos of the preferred sex by selected capacitation treatment and co\u2010culture can be the method of choice in bulls with low IVF yields from sorted sperm.<\/jats:p>","DOI":"10.1111\/rda.13926","type":"journal-article","created":{"date-parts":[[2021,3,14]],"date-time":"2021-03-14T12:21:20Z","timestamp":1615724480000},"page":"864-871","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":4,"title":["Production of sexed bovine embryos in vitro can be improved by selection of sperm treatment and co\u2010culture system"],"prefix":"10.1111","volume":"56","author":[{"given":"Ivona","family":"Travnickova","sequence":"first","affiliation":[{"name":"Department of Genetics and Reproduction Veterinary Research Institute Brno Czech Republic"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Pavlina","family":"Hulinska","sequence":"additional","affiliation":[{"name":"Department of 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